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<Original Articles>Protective Effects of Combined Therapywith A Protease Inhibitor, ONO3307, and A Xanthine OxidaseInhibitor, Allopurinol on Temporary Ischaemic Model ofPancreatitis in Rats
Author(s) HIRANO, TETSUYA; MANABE, TADAO; OHSHIO,KAKUJI; NIO, YOSHINORI
Citation 日本外科宝函 (1992), 61(3): 224-233
Issue Date 1992-05-01
URL http://hdl.handle.net/2433/203742
Right
Type Departmental Bulletin Paper
Textversion publisher
Kyoto University
Arch Jpn Chir 61(3), 224~233, Mお, 1992
Protective Effects of Combined Therapy with A Protease Inhibitor,
ONO 3307, and A Xanthine Oxidase Inhibitor, Allopurinol on
Temporary Ischaemic Model of Pancreatitis in Rats
TETSUYA HIRANO, TADAO MANABE, GAKUJI 0HSHIO and YosHINORI Nxo
First Department of Surgery, Faculty of Medicine, Kyoto University, Kyoto Japan Recieved for Publication, Feb. 13, 1992
Abstract
The protective effect of a new potent protease inhibitor, ONO 3307, in combination with a xan-
thine oxidase 1出 1bitor,allopurinol, waste町 din pancreatico-biliary duct obstruction (PBDO) w油
temporary pancreatic ischemia in rats.
After PBDO with ischemia, we observed hyperamylasemia, pancreatic edema, congestion of
amylase and lysosomal enzyme cathepsin B as well as impaired output of amylase and cathepsin Bin-
to the pancreatic juice and a redistribution of lysosomal enzyme from the lysosomal fraction to the
zymogen fraction. The administration of ONO 3307 plus allopurinol almost completely prevented
the pancreatic injuries induced by PBDO with ischemia.
These results indicate the important roles of temporaηpancreatic ischemia in the pathogenesis
of pancreatic damage and the usefulness of combination therapy with a new potent protease inhibitor
and xanthine oxidase inhibitor in the protection against clinical acute pancreatitis.
Introduction
It has been suggested that many factors, such as pancreatic duct obstruction1ベhypersecretoη
condition3, bile-reflux4・5, and ischaemia6・7 of the pancreas are closely related to the early events in the
pathogenesis of acute pancreatitis. Moreover, redistribution of lysosomal enzyme from the
lysosomal fraction to the zymogen fraction and colocalization oflysosomal enzymes with digestive en-
zymes have been described in the experimental pancreatitis1・2・3・8・9・10・11. This colocalization of
lysosomal enzymes with digestive enzymes has been suggested to play an important role in the
development of pancreatic injury, because lysosomal enzyme cathepsin B can activate tηrpsinogen
12・13・14, and trypsin can activate many other key enzymes in pancreatitis. Furthermore, reports
have accumulated about the protective effects of new potent synthetic protease inhibitors with
relatively low molecular weights on the exocrine pancreas in various models of acute pancreatitis
Key words: Protease inhibitor, Xanthine oxidase inhibitor, Ischaemia, Redistribution oflysosomal enzyme, Cathep-sin B, Pancreatic secretion of lysosomal enzyme.
索引用語 プロテアーゼインヒビター,キサンチンオキシダーゼインヒピター.虚血,ライソゾーム酵素再分布,
カテプンン B,ライソゾーム酵素の醇性分泌.
Present Address: First Department of Surgery, Faculty of Medicine, Kyoto University, 54 Shogoin-Kawaracho, Sakyo-ku, Kyoto 606, Japan.
ISCHAEMIA AND REPERFUSION IN PANCREATITIS 225
1s,16,11,1s,19, and the use of such a new potent protease inhibitor may be useful in the treatment of
clinical pancreatitis.
In ischaemia models of pancreatitis, the ischaemia and reperfusion relationship has been
reported to play an important role in the pathogenesis of pancreatic injury through the formation of
free radicals20・21 It is possible that free radicals form a common pathway of in ury in several forms
of acute pancreatitis, because there have been several reports about the close relationship of free
radicals in the other models ofpancreatitis: caerulein-induced22, and choline-deficient ethionine sup-
plemented diet-induced23‘ These repo口sabout free radicals suggest that free radical scavengers may
be useful at any time in the treatment of pancreatitis.
In this study we evaluated the protective E仔ectof combined therapy with a new potent protease
inhibitor, ONO 3307 (4・sulfamoylphenyl4-guanidinobenzoate methanesulfonate), and allopurinol
against the multifactor-related pancreatic injury induced by pancreatic biliary duct obstruction in "
temporary ischaemia model.
Materials and Methods
Animal preparation
A total of64 male Wistar rats weighing about 350 g (Funahashi Farm, Shizuoka, Japan) were us-
ed in this study. They were kept in air-conditioned (23±3°C) animal quarters and allowed to
become acclimatized to standard laboratory conditions for at least 4 days. Experiments were begun
after a 16・hourfast. The rats divided into the following 4 groups:
a) Pancreatico-biliary duct obstruction (PBDO) group (12 rats)-After a 16-hour fast, general
anesthesia was induced by intraperitoneal pentobarbital injection (25 mg/kg), and catheterization
(V-3 catheter, lnsul Tab, Woburn, MA, U.S.A.) of the superior vena cava was performed
through the right external jugular vein. The catheter was tunneled beneath the skin of the back to
the root of the tail and brought out. Laparotomy was performed through an upper midline inci-
sion, and the pancreatico-b出aryduct (PBD) was occluded by a small metal clip just adjacent to the
duodenum. Just after occlusion of the PBD, secretion (0.2 CU/kg • hr) (Sigma Chemical, St.
Louis, MO, U.S.A.) and caerulein (0.2 μg/kg・hr)(Si胴 aChemical) were infused for 30 minutes
with an infusion pump. At 1 hour after occlusion of the PBD, the metal clip was removed and the
abdomen was closed. Animals were kept in cages for the next experiments.
b) lschaemia group (14 rats)-As in the PBDO group, the pancreatico・bilia町 ductwas ligated, and
secretin and caerulein were infused for 30 minutes. Then pancreatic ischaemia was induced by
ligation of the caeliac and superior mesenteric arteries with the clips for next 30 minutes. After
this 30・minutespancreatic tempora町 ischaemiawith PBDO, clips to the duct and a目erieswere
removed, and the abdomen was closed.
c) Pretreatment group (16 rats)-As in the ischaemia group, rats were prepared with PBDO and
secretin and caerulein stimulation and ishcaemia. Starting 1 hour before and continuing
throughout the experiment a new potent protease inhibitor, ONO 3307, was infused in a dose of 2
mg/kg • hr, and just before ligation of the PBD, allopurinol (Sigma Chemical) was injected in-travenously in a dose of 30 mg/kg, and just after removal of the ligation of the PBD and of the
arteries, allopurinol (20 mg/kg) was injected again. Thereafter, every 2 hours allopurinol was in-
jected intravenously in a dose of 20 mg/kg for 10 hours (5 times). ONO 3307 was donated by
ONO Pharmaceutical Company, Osaka, Japan.
226 日外宝第61巻第3号(平成4年 5月)
d) Control group (10 rats)-As in the PBDO group, catheterization and laparotomy were performed
and 1 hour after gentle manipulation of the pancreatico-biliary duct and a portion of the caeliac
and superior mesenteric arteries, the abdomen was closed. In addition, other two groups; PBDO
with caerulein and secretion stimulation and ischaemia plus only ONO 3307 group (n=6) and on司
ly allopurinol group (n=6) were used.
All the rats received heparinized (30 U/ml) lactate-Ringer solut
the whole巴xperiment(also a丘巴rsurg巴ry)and were kept on heating pads at 40。Cunder overhead
lamps, to maintain core temperature. After surgery, all the animals were fasted but given free ac-
cess to tap water. Twelve hours after the removal of the ligations from the PBD and arteries, rats
were sacrificed by a large does of intravenous pentobarbital.
Serum a九ylaselevels and pancreatic water, amylasιand cathψsin B content
The abdomen was reopened, and blood was drawn from the inferior vena cava for the determina-
tion of serum amylase levels. Portions of pancreas were removed quickly and one small po口ionwas
used for the determination of pancreatic oedema by a comparison of the weight immediately after
sacrifice (wet weight) to that of the same sample after incubation at 150°C for 48 hours in a desiccator
(dry weight). Other small portions of the pancreas were homogenized in 5 ml ice-chilled phosphate-
buffered saline (pH 7.4) containing 0.5% Triton X-100 (Fisher Scientific) in a Brinkmann Polytron
(Brinkmann Instruments, Inc., Westbury, NY, U.S.A.) for the measurement of the pancreatic con-
tent of amylase and cathepsin B, as a lysosomal enzyme. Both amylase and cathepsin B activity, as
well as deoxyribonucleic acid (DNA) concentration, were measured in the resulting supernatant
after low speed centr出 gation(150×g, 15 min, 4°C) and were expressed as U/mg DNA.
Histological changes
Other veηsmall portions of pancreas from each group were fixed overnight in phosphate-
buffered (pH 7.4) 10% neutral formalin solution. After para伍nembedding, sectioning and staining
with hematoxylin-eosin, the sections were examined by a blinded observer, and intestinal edema,
acinar cell vacuolization, inflammatory cell infiltration, and acinar cell necrosis were graded on a
0-4+ scale (0, no change; 4+, maximum change).
Subcellular distribution of amylase and cathepsin B activi汐
The remaining portions of the pancreas were homogenized in 6 ml of ice-chilled 5 mM MOPS
(3・(N-mo叩holino)propanesulfonic acid) (Sigma Chemical) buffer (pH 6.5) containing 1 mM
MgS04 and 250 mM sucrose by 3 up-and-down strokes of a Dounce homogenizer (Wheaton,
Millville, NJ, U.S.A.) in a cold room. Unbroken cells and debris were removed by low speed cen-
trifugation (150×g, 15 min, 4°C). The resulting supernatant was considered to be the entire sam-
ple for later calculations and to contain 100% of all measured components. Subcellular fractiona-
tions were performed by differential centrifugation, as described by Tartakoff and J amieson24, with
minor modifications for studies of rat tissue25 Briefly, the supernatant described above was cen-
trifuged (1300×g, 15 min 4°C) to yield a“zymogen granule" pellet and another supernatant. This
supernatant was centrifuged (12,000×g, 12 min, 4°C) to yield a“combined lysosomal and mitochon-
drial”pellet and a supernatant, considered to be the “microsomal and soluble" fraction. The
various pellets obtained during fractionations were resuspended indivudually in 2 ml of ice-chilled 5
mM MOPS buffer, and the amylase and cathepsin B activity in each fraction was measured and ex-
pressed as a percentage of the total activity as an index of distribution of both digestive and lysosomal
enzymes in the pancreatic acinar cells.
In-vivo amylase and cathepsin B ouφut
227
Other new rats were used in this in-vivo experiment. For each group, at selected time, animals
were anesthetized with intravenous pentobarbital (25 mg/kg) and the abdomen was reopened. After
the formation of an external biliary fistula (PE 10, Clay Adams, Parsippany, NJ, U.S.A.) at the liver
hilum, the pancreatico-biliary duct was cannulated (PE 50, Clay Adams) just adjacent to the
duodenum. After 30 minutes for stabilization, caerulein was infused in a dose of0.2 μg/kg • hr for 1
hour for stimulation of pancreatic secretion, and pancreatic juice was collected in pre-weighed eppen-
dorf tubes on ice. The volume of pancreatic juice, and amylase and cathepsin B activities were
measured. Both amylase and cathepsiin B outputs were expressed as U/kg • hr.
Assays
Amylase activity was measured by the method of Bernfeld26 with soluble starch as the substrate,
and cathepsin B activity was measured fluorometrically by the method of McDonald and Ellis27 with
CBZ-arginly-arginine-,B-naphthylamide (Bachem Bioscience, Philadelphia, PA, U.S.A.) as the
substrate. Deoxyribonucleic acid (DNA) concentration was measured fluorometrically by the
method of Labarca and Paigen28, with calf thymus DNA (Sigma Chemical) as the standard.
Statistical analysis
This results reported in this study represent the mean±SEM for n determinations. Differences
between groups were evaluated by ANOBA and Tukey method. For evaluating the histological
changes, Wilcoxon rank-sum test was used and significant differences were defined as those
associated with a probability value (p) ofless than 0.05 (p<0.05).
ISCHAEMIA AND REPERFUSION IN PANCREATITIS
Results
All the animals in the PBDO and ischaemia groups as well as those in the treatment group sur-
vived after surgery.
Serum amylase levels, pancreatic water, amylase and cathepsin B content
One hour pancreatico-biliary duct obstruction (PBDO) with 30 minutes of stimulation by secre-
tion (0.2 CU/kg ・ hr) and caerulein (0.2 μg/kg ・ hr) plus 30 minutes of temporary pancreatic
ischaemia caused a significant increase in serum amylase levels, pancreatic water, amylase and
cathepsin B content, but PBDO with secretion and caerulein stimulation without ischaemia did not
Effects of ONO 3307 and allopurinol on the changes in serum amylase levels, pancreatic water, amylase and cathepsin B con臼ntin rat panロeati co・biliaryduct obstruction (PBDO) with pan口四ticischaemia
Pancreatic amylase content
巴担KQ旦企i573土62*
728±54
481 ±57判
439±45本本牟
592土48本
604±51
Pancreatic cathepsin B content 巴生~剖1226± 138事
1724± 189
1275± 153*
1182±126**
1347±165
1423±154
Table 1
Pancreatic water content
(%of wet weight)
Serum amylase levels (U/ml)
77±2
81±2
76±2*
74±1件
78±2
79±2
8±2料
23±3
9±2**
6± 1 ***
14±2本
16±2
n
ぷU
勺’oo
,コζ
U
ζ
U
Group
PBDO
Ischaemia
Pretreatment
Control
ONO 3307
Allopurinol
PBDO:・ pancreatico-biliaryduct obstruction, Ischaemia:目 PBDOwith pancreatic ischaemia, Pretreatment: PBDO with pancreatic ischaemia plus ONO 3307叩 dallopurinol administration, Control: controlled laparotomy groups, ONO 3307: PBDO with pancreatic ischaemia plus only ONO 3307 group, Allopurinol: PBDO with pancreatic ischaemia plus only allopurinol group, *p<0.05, **p<0.02, and 料*pく0.01compared with ischaemia group
228 日外宝第61巻第3号(平成4年 5月)
Table 2 Effects of ONO 3307四 dallopurinol on the changes in pancreatic acinar cell histology in rat PBDO wnh ischaemia
Histological changes of p阻 creaticacinar cells
Group n Interstitial Acinar cell lnfi副nma torγ Acinarcell cell edema vacuohzat10n
infiltration necrosis
6 0.3±0.2* 0.3±0.2* 0.2±0.2* 0.2±0.2* PBDO (0-1) (0-1) (0-1) (0--1)
7 2.1±0.3 2.4±0.2 2.3土0.3 1.9±0.3 lschaem旧 (1-3) (2-3) (1-3) (1-3)
8 0.4±0.2* 0.3±0.2* 0.3±0.γ 0.2±0.2牟Pretreatment (0--1) (0--1) (0--1) (0--1)
。** 。** 。本* 。**
Control 5 (0) (0) (0) (0)
ONO 3307 6 1.2主0.2 1.2 ±0.2* 1.2土0.4 1.2±0.2 (1-2) (1-2) (0--2) (1-2)
Allopurinol 6 1.4土0.2 1.2±0.2* 1.4±0.4 1.4±0.4 (1-2) (1-2) (1-3) (1-2)
Histological changes were blindedly graded on a scale from 0 (no changes) to 4+(maximum changes and出evalues 訂 eexpressed as means±SEM, ( ); the r釦 gesof批 values. The symbols田町groupsare山 sameas in Table I. う<0.05, 叫 pく0.02compared with ischaemia group by Wilcoxon rank-sum test.
cause significant changes in these parameters. The administration of ONO 3307 and allopurinol
had a significant protective effect against the hyperaylasemia, pancreatic oedema, and congestion of
pancreatic digestive and Iysosomal enzyme in acinar cells induced by PBDO with hypersecretion
plus temporary pancreatic ischaemia. The administration of only ONO 3307 also had a significant
protective effect against the hyperamylasemia and congestion of pancreatic amylase, but less than the
combination administration. The administration of only allopurinol also had a significant protec-
tive e仔ectagainst the h沖 eramylasemia,but less than the combined administration (Table 1).
Histological chaπ>.ges
PBDO with hypersecretion plus tempora円 pancreaticischaemia caused moderate but signifi-
cant histological changes: interstitial oedema, acinar cell vacuolization, inflammatory cell infiltra-
tion, and acinar cell necrosis. PBDO with hypersecretion induced by secretion and caerulein
without ischaemia caused only very mild histological changes. The administration of ONO 3307 in
combination with allopurinol almost completely prevented the acinar cell damage induced by PBDO
with ischaemia. The administration of only ONO 3307 or allopurinol also had a significant protec-
tive effect against the h国ologicalchanges only in the acinar cell vacuolization (Table 2).
Subcellular distribution of amylase and cathepsin B activity
One hour of PBDO with 30 minutes of stimulation with secretion and caerulein plus 30 minutes
of ischaemia caused a significant decrease of amylase activity in the z戸nogengranule fraction叩 da
significant increase in the microsomal and soluble fraction. PBDO without ischaemia caused no
significant changes. The administration of ONO 3307 in combination with allopurinol had a si伊ifi・
cant protective effect against the increased fragility of zymogen granules induc巴dby PBDO with
ischaemia. PBDO with ischaemia also significantly increased cathepsin B activity in zyrnogen
granules and significantly decreased it in the lysosomal fraction, indicating a redistribution of
lysosomal enzyme from the lysosomal to the heavier zymogen fraction and a colocalization of
lysosomal enzyme with pancreatic digestive enzyme. PBDO without ischaemia caused no signifi-
cant changes in cathepsin B activity distribution. The administration of ONO 3307 in combination
229 ISCHAE恥UAAND REPERFUSION IN PANCREA TITIS
Effects of ONO 3307 and allopurinol on the changes in pancreatic subcellular distribution of amylase and cathepsin B activity in rat PBDO with ischaemia
Table 3
Fractions (% of total)
Mycrosomal and soluble
44±2*
53±3
46土2*
42±2**
47±3
49土2
Lysosomal
17±2
16土2
17±2
18±2
18±2
17±2
Zymogen
39±2判
31±2
37土2*
40±2**
35±5
34±2
PO句
J
Q
U
民
Jroro
Group
PBDO
Ischaemia
Pretreatment
Control
ONO 3307
Allopurinol
Amylase activity
20±2
22土2
18±2
17±2
19±2
20±2
57土2材
40±3
54±2材
59±2料*
50±2
46±3
PBDO
Ischaemia
Pretreatment
Control
ONO 3307
Allopurinol
*p<0.05,時p<0.02,and件*p<0.01compared m由 ischaemiagroup
27±2料
38±3
28±2***
24±2仲キ
31土2
34±3
6
7
8
5
6
6
Cathepsin B activity
with allopurinol significantly inhibited the redistribution of cathepsin B in acinar cell induced by
PBDO with ischaemia. The administration of only ONO 3307 or allopurinol had no significant
changes in the distribution of cathepsin B and amylase in acinar cells (Table 3).
In-vivo amylase and cathepsin B output
In the ischaemia group pancreatic exocrine functions stimulated by caerulein (0.2 μg/kg • hr)
were significantly lower than in the control group. Pancreatic juice volume after caerulein stimula-
tion was significantly lower than in the control group. PBDO without ischaemia caused only a
slight, but not-significant reduction. The administration of ONO 3307 with allopurinol had a
significantly protective effect against the reduction in pancreatic juice volume induced by PBDO
with ischaemia. Both amylase and cathepsin B outputs showed similar changes, and 12 hours after
PBDO with ischaemia, both amylase and cathepsin B outputs after caerulein stimulation were
significantly lower than in the control group. PBDO without ischaemia only a slight, not-significant
decrease in amylase and cathepsin B output. Administration of ONO 3307 in combination with
allopurinol significantly improved both amylase and cathepsin B outputs after caerulein stimulation
(Table 4).
Effects of ONO 3307 and allopurinol on the changes in caerulein induced pancreatic juice volume, amylase
and cathepsin B output in rat PBDO with ischaemia
Table 4
Cathepsin B output
巴生亙~20±2本
7土3
18±3*
23土2**
Amylase output
巴生互生}7335±843*
2889±328
7127土946*
8724±921 **
Juice volume 但些豆回1.58±0.21 *
0.61土0.17
1.48±0.23*
1.72±0.19糾
n
FO
勺
j
o
O
民J
Group
PBDO
Ischaemia
Pretreatment
Control
*p<0.02叩 d件 p<0.01compared with ischaemia group
230 日:外宝第61巻第3号(平成4年5月)
Discussion
In this present study, we evaluated the effects of short-term pancreatico-biliary duct obstruction
and stimulation with secretion and caerulein plus temporary pancreatic ischaemia on the exocrine
pancreas in rats, and the protective effect of a new potent synthetic protease inhibitor, ONO 3307, in
combination with a well-known xanthine oxidase inhibitor, allopurinol. Short-term PBDO without
ischaemia did not cause significant changes in any of the parameters examined in this study, but if
ischaemia was superimposed on PBDO, pancreatic exocrine functions were significantly and marked-
ly impaired. Short-term PBDO alone, or ischaemia alone also caused no significant changes in any
of the parameters (data not shown). Moreover, PBDO with ischaemia caused a marked redistribu-
tion oflysosomal enzyme from the lysosomal fraction to the heavier zymogen fraction, indicating col-
ocalization of lysosomal enzymes with digestive enzymes in the same subcellular compa口mentin
acinar cells. This redistribution oflysosomal enzymes has also been described in other experimental
models of pancreatitis: caerulein-induced8ペpancreaticduct obstruction 1-3 and diet-induced10.u
Thus, this redistribution phenomenon seems to be an important triggering event in the development
of acute pancreatitis. In a recent study, we noted the secretion oflysosomal enzymes into pancreatic
juice stimulated by pancreatic secretagogues2・29 This secretion oflysosomal enzyme into pancreatic
juice seems to be closely related to changes in the transport and distribution of lysosomal enzyme in
pancreatic acinar cells. PBDO with ischaemia also caused marked impairment of both cathepsin B
and amylase secretion during stimulation by caerulein which suggests a disturbance in the sorting of
lysosomal enzymes in acinar cells.
Pancreatic ischaemia, such as that caused by induced hypotension, has been reported to affect
the severity of pancreatitis30, but in the ischaemia models of pancreatitis, the production of oxygen-in-
duced free radicals seems to be more responsible for the injury produced by ischaemia-reperfusion.
In the ischaemia-reperfusion pancreatitis models, xanthine oxidase has been reported to be the
source of superoxide radicals30・31・32・33, because xanthine oxidase is a ubiquitous enzyme in the
gastrointestinal tract and the pancreas also has considerable amounts of this enzyme.
もγetested the protective effect of allopurinol, a specific xanthine oxidase inhibitor, in the
ischaemia-reperfusion model of pancreatitis superimposed on short-term pancreatic duct obstruc-
tion. ONO 3307 is a guanidino acid ester. Such new potent synthetic protease inhibitors have been
re po口edto inhibit strongly many key pancreatic enzymes, such as trypsin, elastase, phospholipase A
2, thrombin, and kallikrein33・H Moreover, its relatively low molecular weight (430 daltons) should
help it to penetrate into the acinar cells, so we were encouraged to evaluate its protective effects in
this models. As in other previous studies and our own recent study15,l6,l7,IB,l9, ONO 3307 showed
protective e百ectsagainst pancreatic injury. In this ischaemia-reperfusion model we tested it in com-
bination with the xanthine oxidase inhibitor, allopurinol.
Our present study strongly suggests that temporary pancreatic ischaemia and oxygen-derived
free radicals generated by xanthine oxidase play crucial roles in the pathogenesis and development of
acute pancreatitis, and ifischaemia is superimposed on pancreatico・biliaηductobstruction, the pan-
creatic damage would be expected to be more severe. The present study also suggest that this new
potent synthetic protease inhibitor in combination with the xanthine oxidase inhibitor, allopurinol,
might be of use in the protection for clinical pancreatitis, although, in this study, the agents were ad-
ministered before the induction of acute pancreatitis rather than subsequently.
Finally, because many factors, such as pancreatico-biliary duct obstruction, intraductal
ISCHAEお1IAAND REPERFUSION IN PANCREATITIS 231
hypertension, and pancreatic ischaemia seem to be closely related to the pathophysiology of pan-
creatitis, this present model of PBDO with ischaemia seems to be an aid and of benefit in the clarifica-
tion of the early events of the pathogenesis of acute pancreatitis as well as possible mechanisms and
the protective effects of various agents, because in its several aspects it represents moderately severe
pancreatit1s.
Acknowledg皿 ents
This study was supported by a grant, Scientific Research B-03454319, from the Ministry of Education, Science and
Culture, and a grant from the Ministry of Health and Welfare of Japan. The authorss thank Mr. Kimiko Hirano for
preparing the manuscript and Ms. Yoko Manabe for typing the manuscript.
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ISCHAE恥11AAND REPERFUSION IN PANCREA TITIS 233
和文抄録
ラットでの短期間総醇胆管結室長,醇管内圧上昇と
醇虚血再還流モデルにおける xanthineoxidase
inhibitorとproteaseinhibitorの併用療法
京都大学医学部第一外科教室
平野鉄也,真辺忠夫,大塩学市,仁尾義則
ラy トにて短期間( 1時間)の総勝胆管結数,
caerulein (0.2 μg/kg・hr)と secretin(0.2 CU/kg・ hr)に
よる勝外分泌刺激および腹腔動脈と上腸間膜動脈結数
(30分後再逮流)による醇炎モデノレにて xanthineox-
idase inhibitorである allopurinolと proteaseinhibitor
である ONO3307の併用療法を検討した.上記モデ
ルにて,高 amylase血症, S革浮腫,解消化酵素の解内
うっ滞,際腺房細胞の空胞化等の組織学的変化,さら
に,細胞分画法にては,隣ライソゾーム膨素のライソ
ゾーム分画よりチモーゲン分画への移動・再分布が観
察されたーしかし, allopurinolとONO3307の併用療
法にてこれらすべての parameterで有意な醇保護効果
が観察されたが, allopurinolとON03307の単独療法
では一部の parameterにのみ保護効果を示したのみで、
あった.これらの結果は,本モデルにおいては, xan-
thine o幻daseを介した oxygen-derivedfree radicalや何
らかの proteaseactivityが,その弊損傷の発生に重要
な役割を果していることとともに,臨床上での弊炎治
療においても,その作用機序が異なった種々の agent
の併用療法の有用性をも示唆させるものであった.
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